Normal spindle morphologies and reduced CENP-E staining at the kinetochores of aligned chromosomes in PIASγ-depleted cells.

<div><p>(<i>A,G</i>) Mitotic spindles (stained with anti-alpha-tubulin antibody) were indistinguishable in control and PIASγ-depleted metaphase cells, except that a small % at late time-points possessed extra poles (arrow in <i>A</i>): (<i>A,G</i>) Red = alpha-tubulin, Blue = DAPI.</p> <p>(<i>B–F,H–K</i>) CENP-E staining (Red) is detected strongly on kinetochores of chromosomes away from the metaphase plate in both control-treated and PIASγ-depleted cells (<i>B,H,I</i>), but was much reduced in chromosomes at the plate (<i>C,J,K</i>).</p> <p>After prolonged arrest in metaphase some of PIASγ-depleted cells had one or two chromosomes that left the plate and regained strong CENP-E staining at their kinetochores (<i>D–F</i> and see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000053#pone.0000053.s002" target="_blank">Fig. S1<i>N,O</i></a>).</p> <p>The same cell is shown in <i>D</i> (merged image), <i>E</i> (CENP-E straining) and <i>F</i> (DAPI straining).</p>